Project Summary/Abstract: Protein-based therapeutics have contributed immensely to health care and constitute a large and growing percentage of the total pharmaceutical drugs. The majority of the biopharmaceutical products are currently manufactured using mammalian cell lines. However, compared to microbial systems, productivity and product titers in mammalian cell cultures are low mainly due to the low biomass concentration achievable in standard mammalian cell culture. In addition, mammalian cell lines produce significant amounts of waste products such as lactate, alanine, and ammonia, which reduce biomass yield and protein production, cause toxic accommodation, and inhibit cell growth. Currently, media and process optimization strategies in mammalian cell culture are performed using a trial and error approach where process outputs are improved laboriously by experimentation. These empirical optimization techniques are widely used because in most cases little is known about the underlying physiological interactions that impact growth and protein production in the host cell lines. A fundamental understanding of cell line physiology and metabolism can greatly improve and accelerate media and process development in mammalian cell line systems. A rational design approach in media optimization and process development requires modeling and simulation technologies capable of capturing and analyzing the underlying physiology of the host cell line. In this SBIR research plan, we intend to demonstrate the value of metabolic modeling in media optimization using a reconstructed model of murine hybridoma cell line. In combination with our modeling technology and using the reconstructed model of hybridoma metabolism, we will design in silico-derived media formulations that reduce byproduct formation in hybridoma cell culture and subsequently evaluate our media designs using experimental measurements. Success of this proposal will demonstrate the scientific and technical feasibility of model-driven improvements in recombinant protein production through the rational selection of nutrient supplementation strategies and ultimately reduce the cost of therapeutic protein development and manufacturing to ensure that the next generation of medicines can be created in amounts large enough to meet patients' needs, and at a price low enough that patients can afford. Project Narrative: Protein-based therapeutics have contributed immensely to health care and constitute a large and growing percentage of the total pharmaceutical drugs. Success of this proposal will demonstrate the scientific and technical feasibility of model-driven improvements in recombinant protein production through the rational selection of nutrient supplementation strategies and ultimately reduce the cost of therapeutic protein development and manufacturing to ensure that the next generation of medicines can be created in amounts large enough to meet patients' needs, and at a price low enough that patients can afford. [unreadable] [unreadable] [unreadable]